基于DNA条形码的甘草属植物的分子鉴别  

Molecular Identification in Genus of Glycyrrhiza Based on DNA Barcoding

作　　者：李珍珍 郑司浩 尚兴朴 邓庭伟 王继永 赵润怀 LI Zhen-zhen;ZHENG Si-hao;SHANG Xing-pu;DENG Ting-wei;WANG Ji-yong;ZHAO Run-huai(Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,China;China National Traditional Chinese Medicine Co.,Ltd,Beijing 100195,China)

机构地区：[1]江西中医药大学,江西南昌330004 [2]中国中药有限公司,北京100195

出　　处：《中国现代中药》2020年第2期207-212,218,共7页Modern Chinese Medicine

基　　金：国家中药标准化项目(ZYBZH-Y-ZY-45);国家重点研发计划(SQ2017YFC170424);国家中药材产业技术体系遗传改良研究室建设(CARS-21);北京市科技计划课题(Z171100001717028);中医药公益性行业科研专项(201507002-1-10)。

摘　　要：目的:评价不同DNA条形码序列对甘草属的鉴定能力。方法:应用5条常用DNA条形码序列ITS、ITS2、psbA-trnH、matK与rbcL对甘草属进行PCR扩增与序列测序,结合在GenBank数据库下载的序列数据,通过分析各序列种内与种间变异与遗传距离,构建系统发育树,并基于相似性搜索算法和最近距离法计算各序列鉴定成功率,以分析比较各序列对甘草属的鉴定效率。结果:matk序列因扩增与测序成功率均较低,没有加入后续的数据分析。各个序列的鉴定成功率由高到低的顺序为psbA-trnH、ITS、ITS2、rbcL。ITS和ITS2序列种间最小变异均大于种内最大变异,且种内变异最小。Barcoding gap检验结果显示,ITS、ITS2的重叠区较小。NJ树结果显示,只有psbA-trnH序列能把甘草属不同物种分开。结论:利用DNA条形码序列可准确鉴别甘草属植物,psbA-trnH和ITS组合序列可作为鉴定甘草属植物的较优序列组合。本研究为甘草属植物的分子鉴别提供科学依据。Objective:To evaluate the identification ability of different DNA barcodes for the genus of Glycyrrhiza.Methods:Five DNA barcodes of ITS,ITS2,psbA-trnH,matK and rbcL were amplified and sequenced,coupled with the sequences downloaded from the GenBank database to analyze the genus of Glycyrrhiza in term of the variation of inter-and intra-species,barcoding gap,and identification efficiency which is evaluated by Nearest distance and BLAST1 methods.The neighbor joining tree(NJ tree)for each barcode were constructed.Results:The order of success rate of identification of each sequence from high to low was as following psbA-trnH,ITS,ITS2,rbcL.The minimum inter-specific distance of ITS and ITS2 sequences was greater than the maximum intra-specific distance,and the minimum intra-specific distance.Barcoding gap test results showed that the overlap area between ITS and ITS2 was the least.The results of NJ tree showed that only psbA-trnH sequence could distinguish different Glycyrrhiza.Conclusion:DNA barcoding could identify the plants of Glycyrrhiza accurately.The psbA-trnH and ITS sequence could be used as an ideal combination for identification of Glycyrrhiza.This study should provide scientific basis for molecular identification on the genus of Glycyrrhiza.

关 键 词：甘草属 DNA条形码 分子鉴别 内转录间隔区 

分 类 号：R282[医药卫生—中药学] S567.71[医药卫生—中医学]